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. 2014 Dec 23;9(12):e116074. doi: 10.1371/journal.pone.0116074

Figure 6. α4 facilitates formation of PP2A AC core enzyme.

Figure 6

(A) Equal amounts of BiFC expression constructs encoding YN-Aα and PP2Acα-YC in the presence of equal amounts of pcDNA5/To-Flag-α4WT, pcDNA5/To-Flag-α4MUT, or empty vector were transfected into NIH3T3 cells. YFP signals due to BiFC of YN-Aα and PP2Acα-YC were measured by fluorescence microscopy. Expression of Flag-α4WT or Flag-α4MUT was verified using anti-FLAG antibody and Cy3-conjugated secondary antibody. DAPI was applied for staining of nuclei. Scale bars, 20 µm. (B) BiFC expression constructs encoding YN-Aα and HA-PP2Acα-YC and pcDNA5/To-Flag-α4WT, pcDNA5/To-Flag-α4MUT, or empty vector were transfected into NIH3T3 cells. Cell lysates were collected 24 h post-transfection, and immunoprecipitations were performed using anti-HA Sepharose. The cell lysates and anti-HA immunocomplexes were then analyzed by SDS-PAGE and Western blotting using the indicated antibodies.